Objective To explore the effect of Mycobacterium tuberculosis antigen (Mtb-Ag) on neutrophils apoptosis.Methods The fresh isolated neutrophils from healthy adults blood were cultured with Mtb-Ag for 24 h,with or without pretreatment of nuclear factor -?B (NF-?B) inhibitor N-tosyl-L-phenylanyl chloromethyl ketone (TPCK) for 30 minutes.Annexin V staining and Flow cytometry were used to measure cell apoptosis of neutrophils.NF-?B DNA binding was measured by gelelectrophorestic mobility shift assay (EMSA) in neutrophils after incubated with Mtb-Ag for 0,1,2,4,6,24 hours.Results Comparing to the spontaneous apoptosis (55%?6%) of neutrophils after culture in vitro for 24 h,treatment of Mtb-Ag (1.125 mg/ml) decreased the cell apoptosis of neutrophils (32%?3%).The NF-?B shift bands were detected at 1 h in neutrophils after stimulated by Mtb-Ag,and reached maximum peak at 2 hours,and then returned to basal levels within 24 h.Pretreatment of TPCK inhibited the anti-apoptosis role of Mtb-Ag in neutrophils.Conclusion Mtb-Ag prevents neutrophils apoptosis and its inhibitory role concerns NF-?B pathway.

BACKGROUND:Titanium and titanium aloy are used mostly in artificial joints, fracture fixation, and oral transplantation, while there are complex cases of insufficient bone mass in these areas. The deepened research of stem cels offers a solution for bone injury to promote new bone formation. The biocompatibility of titanium and stem cels and optimization of titanium surface modification have aroused people's attention. OBJECTIVE:To investigate whether the biocompatibility of titanium and human adipose-derived mesenchymal stem cels can be improved by type I colagen modification of titanium sheets. METHODS:The experiment was divided into two groups. Modification group: titanium sheet was modified with type I colagen; control group: titanium sheet was not modified with type I colagen. Human adipose-derived mesenchymal stem cels at passage 6 were implanted into titanium sheet in two groups. Then we calculated the number of adherent cels in two groups at 1, 2 and 4 hours after implantation, and compared the celladhesion rate. MTT assay was used to observe the proliferation of cels on titanium sheet at 2, 4, 6 and 8 days after implantation. DNA and protein content of cels were detected at 3, 6, 9 days after implantation. The growth of human adipose-derived mesenchymal stem cels seeded upon the titanium sheets was observed under scanning electron microscope at 6 days. RESULTS AND CONCLUSION:When the cels were cultured for 1 hour and 2 hours, the number of adherent cels in the modification group was higher than in the control group (P < 0.05). The absorbance of cels in two groups was increased as the culture time, as detected by MTT assay. The modification group had a significantly higher absorbance value than the control group at 4, 6, 8 days (P < 0.05). DNA and protein contents of the cels in the modification group were higher than that in control group at 6 and 9 days (P < 0.05). At 6 days, the number of adherent cels and secretion of adherent stromal cellmatrix in the modification group were significantly better than that in control group, observed by scanning electron microscopy. Type I colagen modified titanium sheets have good surface activity and biocompatibility, and can promote the proliferation of human adipose-derived mesenchymal stem cels.

BACKGROUND:The safety and efficacy of simultaneous bilateral total knee replacement or selective unilateral total knee arthroplasty in patients with severe osteoarthritis of the knees are stil controversial. OBJECTIVE:To compare safety and clinical efficacy of patients with osteoarthritis knees after simultaneous bilateral total knee replacement or selective unilateral total knee replacement. METHODS:Total y 60 cases with severe osteoarthritis of the knees (90 knees) undergoing total knee replacement were divided into unilateral total knee replacement group (n=30, 30 knees), and the simultaneous bilateral total knee replacement group (n=30, 60 knees). RESULTS AND CONCLUSION:There was no significant difference in the incidence of other complications such as infection, mortality, pulmonary embolism in patients of both groups (P>0.05). The incidence of cardiovascular complications, postoperative blood loss and blood transfusion were higher in the bilateral knee group than in the unilateral knee group (P<0.05). During fol ow-up at 1 year after replacement, no significant differences in range of motion, muscle strength of quadriceps and hospital for special surgery knee score were detected in patients of both groups (P>0.05). However, Visual Analogue Scale scores were significantly lower in the bilateral knee group than in the unilateral group (P<0.05). These data indicated that the risk of cardiovascular complications was high in patients receiving bilateral total knee replacement. Patients with severe cardiovascular disease should avoid simultaneous bilateral total knee arthroplasty.

Aged ; Bronchoscopy ; Esophageal Neoplasms ; complications ; Female ; Humans ; Male ; Middle Aged ; Stents ; Thyroid Neoplasms ; complications ; Tracheal Stenosis ; etiology ; therapy

OBJECTIVESTo detect whether there is an expression of human augmenter of liver regeneration (hALR) in HepG2 cells. To develop a kind of RNAi that specifically targets human augmenter of liver regeneration by synthesizing small interfering RNA (siRNA) in vivo, and to assess the inhibitory effect of this siRNA on hALR expression.

METHODSThe expression of hALR in HepG2 cells was observed with immunocytochemistry. The RNAi plasmid pSIALR-A and the unrelated control plasmid pSIALR-B were transfected into HepG2 cells. Forty-eight hours after transfection, the protein level of hALR was measured with immunocytochemistry; meanwhile, the reverse transcription PCR (RT-PCR) was performed to detect the expression of hALR mRNA.

RESULTShALR was expressed by HepG2 cells. siRNA plasmid pSIALR-A, which targets the cDNA of hALR and the unrelated control plasmid pSIALR-B, was successfully constructed. Both immunocytochemistry and RT-PCR showed that pSIALR-A inhibited the expression of hALR in HepG2 cells significantly, compared with that of pSIALR-B.

CONCLUSIONThe results showed that the small interfering RNA targeting hALR suppresses the expression of hALR in a sequence-specific manner

Base Sequence ; Humans ; Liver Neoplasms ; genetics ; metabolism ; pathology ; Molecular Sequence Data ; Plasmids ; genetics ; Proteins ; genetics ; pharmacology ; RNA, Small Interfering ; genetics ; Transfection

BACKGROUNDWhether WW domain containing oxidoreductase (WWOX) gene is a tumor-suppressor is still controversial. Some researchers found that the transcription of the WWOX gene was lacking not only in tumor tissues but also in non-tumorous tissues and sometimes in normal tissues. Hence it is important to explore the role of the expression of the exogenous WWOX gene in the proliferation and apoptosis of primary cultured lung carcinoma cells.

METHODSLipofection technique was used to determine primary cultured lung carcinoma cells containing the highly expressed exogenous WWOX gene and primary cultured cells with vectors as controls. An animal model of lung cancer was made by subcutaneous implantation of tumor cells into nude mice. RT-PCR, Western blotting, flow cytometry, and TUNEL were used to detect the transcription, expression of the exogenous gene and the effect of the expression of targeted genes on the proliferation and apoptosis of the primary cultured lung carcinoma cells.

RESULTSThe growth, clone formation rate (CFR) ((5.33 +/- 1.53)%) of the primary lung cancer cells transfected with the WWOX gene, tumor size and weight were significantly lower than those of the non-transfected lung cancer cells (CFR: (14.33 +/- 1.53)%) and the primary lung cancer cells transfected with blank plasmids (CFR: (11.00 +/- 1.73)%, P < 0.05). The apoptosis level of primary lung cancer cells transfected with the WWOX gene ((40.72 +/- 5.20)%) was significantly higher than that of the non-transfected lung cancer cells ((2.76 +/- 0.02)%) and the primary lung cancer cells transfected with blank plasmids ((2.72 +/- 0.15)%, P < 0.05).

CONCLUSIONThe expression of the exogenous WWOX gene can significantly inhibit the proliferation of lung cancer cells and induce their apoptosis, suggesting that the WWOX gene possesses tumor-suppressing effect.

Animals ; Apoptosis ; Carcinoma ; pathology ; Cell Line, Tumor ; Cell Proliferation ; Humans ; Lung Neoplasms ; pathology ; Mice ; Mice, Inbred BALB C ; Oxidoreductases ; genetics ; physiology ; Phenotype ; Tumor Suppressor Proteins ; genetics ; physiology ; WW Domain-Containing Oxidoreductase

OBJECTIVESTo examine the activities of transcription factors (TFs) in human hepatocellular carcinoma (HCC) cell lines with different metastatic potentials, so as to identify the TFs associated with HCC metastasis.

METHODSTranscription factor activity profile of Hep3B, MHCC97L and MHCC97H, three HCC cell lines with different metastatic potentials, were examined using protein/DNA array. Electrophoretic mobility shift assays (EMSA) and Western blot were used to confirm the results obtained by protein/DNA array.

RESULTSFrom a total of 345 screened TFs, 7 activity differential TFs were found, of which 5 showed increased activity, including p53, hypoxia inducible factor-1 alpha (HIF-1alpha), signal transducer and activator of transcription 3 (Stat3) and Sp1, and 2 showed decreased activity including Rb and Smad3.

CONCLUSIONThe abnormal functioning of transcription factors is closely associated with HCC metastasis. Our present findings could be of help in expanding our understanding of the mechanism of HCC metastasis and identify new predictive biomarkers and therapeutic targets.

Carcinoma, Hepatocellular ; metabolism ; pathology ; Cell Line, Tumor ; DNA Fingerprinting ; Humans ; Liver Neoplasms ; metabolism ; pathology ; Neoplasm Metastasis ; Protein Array Analysis ; Transcription Factors ; classification ; genetics ; metabolism

Plasma obestatin level was determined in patients with impaired glucose regulation and type 2 diabetes mellitus.The plasma obestatin levels in patients of both groups were significantly decreased as compared with that in controls.Plasma obestatin level was negatively correlated with body mass index,HbA_(1C),waist-to-hip ratio,plasma insulin and HOMA-IR.Obestatin level seems to be related with metabolic disorder.

Objective To evaluate the imaging methods,characteristics,diagnostic value of multi- slice CT urography(MSCTU)in congenital abnormities of urinary system.Methods To collect 33 urinary congenital abnormities cases in three years and to analyses these MSCTU images.All examinations were performed with a multi-slice spiral CT scanner.The patients were intravenously injected with 90 ml of Iohexol 300 with a power injector at the rate of 3 ml/s.Nephrographic-phase images were obtained at 75 s after initiation of the injection of contrast material,the appropriate delay time is according to Kidney's enhancement extent and nephrohydrosis degree.Excretory-phase images were obtained through the abdomen and pelvis from 10 min.to 23 h after initiation of the injection of contrast material without abdominal compression.Excretory-phase images were transferred to the workstation and performed maximun intensity projection(MIP),multiplanar reconstruction(MPR),volume rendering(VR),and virtual cystoscopy (VC).Results The urinary congenital abnormities diagnosed by MSCTU in 33 cases,including 1 ectopic kidney,1 horseshoe kidney,1 renal malrotaion,2 supernumerary kidneys,2 ureteral valves,2 retrocaval ureters,4 congenital megaureters,6 ureteropelvic junction stenosis,9 pelviureteric duplication malformations and 5 bladder diverticula.The displaying rate of ureter was 91%(61/66).The scanning time of excretory-phase was less than 20 seconds in All cases.The average CT value of contrast media in displayed ureter lumens was 520 HU.The postprocessing images had clear,dimensional feature and It was satisfy the diagnosis.Conclusion MSCTU has clear,dimensional feature and has strong ability of displaying total anatomy shape and tiny pathology change of congenital abnormities in the urinary system.It is a very useful method for detecting the congenital abnormities in the urinary system.

OBJECTIVESTo screen the hALR-interacting protein by phage-displayed technique and identify its biological activities.

METHODSThe specific phage clones that interacted with target protein hALR from a cDNA library of hepatocarcinoma cells were selected using the T7 phage-displayed technique. The acquired cDNA inserts were sequenced and analyzed by bioinformatic tools. The biological activities of the phage-displayed peptide affecting QGY hepatocarcinoma cells were studied using 3H-TdR method.

RESULTSThe cDNA inserts with 212 bp were acquired after 4 rounds of biopanning. They showed 100% homology with citron kinase. The phage-displayed peptide and the peptide combined with hALR affected QGY cells proliferation.

CONCLUSIONShALR-interacting peptide can be specifically screened by phage-displayed technique. Citron kinase that interacted with hALR potentially plays an important role in the proliferation of hepatocarcinoma cells.

Carcinoma, Hepatocellular ; metabolism ; pathology ; Gene Library ; Humans ; Liver Neoplasms ; metabolism ; pathology ; Liver Regeneration ; Peptide Library ; Peptides ; metabolism ; Protein Binding ; Proteins ; genetics ; isolation & purification ; metabolism ; Tumor Cells, Cultured